Pharmaceutical composition based on collagenase microbial origin
(57) Abstract:The invention relates to medicine, in particular to the creation of a new pharmaceutical compositions containing collagenase microbial origin. The invention consists in that the pharmaceutical composition contains collagenase from Streptomyces lavendulae in the calculation 3-10 IU per 100 g of product. In addition, the composition contains antiseptic Miramistin, proxanol is chosen and polyethylene glycol. The technical result is the creation of a new high-performance non-toxic, non-allergenic pharmaceutical composition for healing and cleansing of the wound, burns and other skin damage based on collagenase microbial origin. 1 C.p. f-crystals, 4 PL. The invention relates to medicine, in particular to the creation of a new pharmaceutical compositions containing proteolytic enzymes for cleansing of wounds and other skin lesions, namely microbial collagenase.Known pharmaceutical compositions that promote the healing of wounds and other damage, reduce the number of complications pathologic process and optimization of the regeneration of the skin, containing antioxidants, immunomodulators, peptidases (Cousin M. I., Kostyuchenko is linoma the study of drugs for local treatment of purulent wounds, M., 1989; EN 2005488, 15.01.94, EN 2106877, 20.03.98.; The possibility of enzyme purification and treatment of wounds. // Symposium M. 1977).For the treatment of skin lesions (wounds, burns, frostbite, and so on) use of pharmaceutical compositions containing collagen. Well known drugs such as "Matural", "Colocal", the collagen sponge."Matural" approved Pharmacological Committee, Ministry of health USSR (Reg. N 87/637/3) is a combination drug consisting of collagen derived from gelievogo Spilka cattle and methyluracil, is a fine dry plate white color with a specific smell. "Matural" is used as local funds in the complex treatment of trophic ulcers, bedsores, deep and long-term healing of wounds, superficial burns of the skin. The drug has anti-inflammatory, stimulates reparative processes in wounds, speeding up the growth and maturation of granulation tissue and epithelization. When applying to the wound "Matural" tightly to its surface, absorbs wound, swelling and gradually literoitca, freeing it contains methyluracil.Sponge "Colocal", the Order of the USSR Ministry of health N 1353 from 30.11.1984 year (Registration N 8/353/scota, furatsilin, boric acid, procaine, honored, is a three-layer porous plate yellow and white with low specific smell. "Colocal" is used as a local treatment for thermal burns II-III degree in the initial stage of their epithelization and for the temporary closure of granulating wounds burns IIIB-IV. The drug is also prescribed for the treatment of donor sites skin, leg ulcers, pressure sores, deep wounds. "Colocal" has a stimulating effect on reparative processes in wounds, speeding up the growth of granulation tissue and epithelization, has anti-inflammatory, hemostatic, and anesthetic properties, and prevents plasmopara through the wound surface.The hemostatic sponge of collagen (Registration N 78/860/7) is prepared from a solution of collagen derived from gelievogo Spilka cattle. The composition of the sponge consists of boric acid and furatsilin. It is a dry porous mass of yellow with a faint odor of acetic acid, absorbs liquid, with slight swells. Sponge has hemostatic and antiseptic, stimulates the regeneration of tissues.Sponge collagen to generate what is a plate yellowish-orange color, it has antimicrobial and anti-inflammatory, stimulates regenerative processes, heals wounds, absorbs wound. Collagen sponge with sanguinarine used to treat infected wounds, including festering postoperative purulent wounds in the 1st and 2nd phases of wound healing, open infected fractures, burns and long-term healing of wounds and ulcers."Digimon" (Moscow VNIIMP and the Institute of surgery. A. C. Wisniewski) - bandage elementality antimicrobial based on the solution of collagen, contains eleven, dioxidine, glutaric aldehyde, is a soft porous plate. "Digipan is intended for the treatment and preparation for autodermoplasty burns IIIa, IIIB and IV degree with a complicated course, after removal of the scab treatment flat infected, slow nonhealing wounds, trophic ulcers, bedsores. The coating absorbs 40+3 g/g of body fluids, parovozduhoduvnaja, plastic, provides a prolonged metered output dioksidina in the wound, stimulates reparative processes in terms of infected wounds, the growth of granulation tissue, accelerates regional and islet EPI collagen, having a range of advantages and some disadvantages, in particular each of these drugs can be used only at a certain stage of wound process, sometimes only for serious injuries and other serious damage to the skin and surrounding tissue, does not provide cleansing of the wounds of necrotic tissue.The most well-known drug with broad applications "Solcoseryl" (instructions for use. Special information "Solco", 1990) for wound healing, resulting from cuts, burns and frostbite, not destroys collagen necrotic tissue and requires a complex individual therapeutic approach in use.The disadvantage of most of the currently used drugs, healing of skin damage is due to their narrow focus of the action, inability to function at different stages of the pathological process, but also to playback increasing the injured epithelium and stimulate regeneration of the skin and surrounding tissue. In addition, they cannot achieve optimal cleansing of wounds, because they do not contain collagenase and not degradable collagen in the native state is the property to the breakdown of collagen with the release of free amino acids hydroxyproline. According to the international classification of enzymes collagenase is 188.8.131.52 (3 hydrolase, 3.4.-acting on peptide bond, 3.4.24. - metalloproteinase (containing calcium).There are two types of collagenases:
1. Collagenase produced by some microorganisms. So. Clostridium histolyticum (the causative agent of gas gangrene) produces collagenase that cleave the polypeptide chain of collagen in more than 200 sites. This collagenase that destroy connective tissue barriers of the host body, promotes penetration (invasion) highly pathogenic Clostridium in the body. Itself the bacterium does not contain collagen and is therefore not subjected to the action of collagenase. (Proteolytic enzymes/ Eds. Colowick, S. P., Kaplan N. O./ Methods in enzymology. 1970. V. 14. Part b, p. 861-872; Mashkovsky M. D. // Medicines. M.: Medicine, 1988, 11 o'clock. S. 55
2. Collagenase, which are found in amphibians and mammals in growing or undergoing a metamorphosis tissues. (Adv. Enzymol. Collagenases and elastases // Ed. Nord, F. F. N. Y. Acad. Press. 1961. V. 23, P. 163-264; Nordwing A. collagenolytic enzymes. N. Y. Acad. Press. 1971. P. 155-206). First collagenolytic activity was detected in the caudal fin tadpoles during metamorphosis, when in a few days is resorption of large quantities of which were cultured explants of skin tissue and the caudal fin tadpoles. After cleaning, the product has been obtained enzyme containing a small number (5%) admixture of other proteases having caseinolytic activity. Collagenase animal tissue has a lower molecular weight than bacterial and differs from the latter by a number of other properties. It was found that the enzyme is inactivated by cysteine and after heating at 50-60owithin 10 minutes is the optimum action at a pH of 7.0 to 8.0.Collagenase is a unique enzyme that is able to selectively hydrolyze insoluble protein collagen, which is part of the connective tissue of a living organism. Splitting is extremely resistant to the action of other proteases collagen fibers with the formation of high molecular weight fragments, collagenase performs in humans and animals with important functions, participates in the transformation of the connective tissue during growth and morphogenesis, as well as to address some of the pathological processes in arthritis, tumor metastasis, etc.Collagenase acting on the collagens of different types isolated from tissues of vertebrates, crabs, larvae of flies, excreta of insects, and microscopic bacteria, fungi and actinomycetes.Ist used in fundamental scientific research, as well as in biotechnology, when necessary efficient, specific and rapid hydrolysis of native and denaturirovannykh collagen fibers of different types. An important field of application is in the treatment of hernia intervertebral disc of the spine, burn wounds, frostbite to accelerate the rejection of scabs and necrotic ulcers, for cleansing of necrotic strikes. In addition, they are required to obtain and cultivation of cell cultures of human and animal tissues, which is the source of activators plasminogen in thrombolytic therapy. Allocated using microbial collagenases viable cells of the islets of Langerhans used in the new method of treatment of diabetes (C. I. Mazurov Biochemistry of collagen proteins. - M.: Medicine, 1974).Among the collagenases microbial origin of the most studied and well known are collagenase synthesized by the bacterium Clostridium histolyticum. Clostridial collagenase are characterized by high specificity of action on collagen fibers and can be classified as "true" collagenases. They are widely used in biotechnology for laboratory purposes. However, for the cultivation of Cl. histolyticum and mass production of collagenase Neveu gangrene and producing toxins. For optimal biosynthesis of collagenases culture Cl. Histolyticum was used complex nutrient medium.Purified clostridial collagenase show activity against native and denatured collagen and act on other proteins, while the crude preparations containing and other proteases. able to cleave not only collagen, and other extracellular material, and therefore are widely used for isolation of cells in various types of animal tissues. As collagenase amphibians, peeled clostridial collagenase can hydrolyze collagen for its complete dissolution, acting as fibrous native collagen and transferline tropocollagen particles.Purified collagenase from Achromobacter iophagus showed a high specificity of action, gidrolizu only collagen and is not acting on other substrates. The activity was 6 times higher than the activity of clostridiales. Properties of collagenases from A. iophagus and Cl. histolyticum was similar. Their activity was not affected by parachlorometaxylenol. Found that both drugs are metallovedenie containing Zn in the active centre and retains activity in the presence of ionic low-molecular collagenolytic proteases new serine-type different from the "true" collagenases primarily by the structure of the active centre. The same collagenase isolated from insect Hypoderma lineatum and crab Uca pugilator.In recent years there have been wound healing compositions containing collagenase.In particular, patent RU 2033805, 30.04.95 describes a method of obtaining a porous wound material containing collagenase solution. The microbial polysaccharide mannan is dissolved in water to a concentration of 6-8 wt.%, and then mixed with an equal volume of collagenase. This medicine performs only one function - cleaning the wound of necrotic tissue does not possess antiseptic properties, does not relieve the wounds from pus and exudate.Closest to the proposed composition is the drug "Iruksol" (Germany, the firm knoll AG D-67008 Ludwigshafen) (Mashkovsky M. D. Medicines. - M.: Medicine, 1988), which contains collagenase in a mixture with a protease isolated from a bacterial culture of Clostridium histolyticum (Handbook "Sigma", 2000, S. 274).1 g ointment contains clostridiales A 0.6 U, chloramphenicol 10 mg related enzymes in anhydrous lipophilic ointment base."Iruksol" is to the WHI - a proteolytic enzyme isolated from Clostridium histolyticum, lysis of necrotic tissue, thereby enzymatic, painless cleansing wounds. In addition, collagenase stimulates granulation process and does not inhibit epithelialization. Collagenase no effect on the intact epithelium, granulation, fat and muscle tissue. Chloramphenicol is a broad-spectrum antibiotic, active against gram-negative and gram-positive bacteria. Cleansing ointment "Iruksol" on the wound in 97% of patients starts with 1 day or no later than the 14th day, while more than 50% of patients effect of the ointment is already apparent within the first 6 days."Iruksol" is used for enzymatic cleansing of wounds, including varicose ulcers, bedsores, gangrene of the extremities, especially in patients with diabetes, as well as for frostbite, nonhealing postoperative wounds, after radiotherapy, in preparation for the facial skin.The lack of similar "Iruxol" is the content in its composition of the antibiotic chloramphenicol, which can cause an allergic reaction and now little effect on not adapted to Iruxol" collagenase derived from pathogenic anaerobic strains of bacteria, and the selection is made using a complex, expensive technologies (Susagawara R., Harper E. Publication and characterization of three forms of collagenase from Clostridium histolyticum // Biochemistry. 1984. V. 23. P. 5175-5182).The technical purpose of this invention is the creation of a new, more efficient and cost-effective wound healing composition which is characterized by a comprehensive course of action, versatility, ability to remove necrotic tissue, pus and exudate from wounds, to carry out rapid healing, to dry up the tissue, to actively remove the scabs, to improve the qualitative composition of scar tissue, to optimize the processes of reparation, to suppress pathogens without causing allergic reactions, and thereby create the possibility of using one product for multidirectional influence on the wound healing process in different stages of its course.This goal is achieved by the fact that a new pharmaceutical composition contains collagenase from Streptomyces lavendulae based 3-10 IU per 100 g of product.The composition may contain auxiliary substances, such as antiseptics, various topical excipients and pharmaceutically acceptable additives. In particular we have studied R> Proxanol is chosen - 22,0 - 30,0 g
The glycol - the Rest is up to 100 g
As the active agent composition selected collagenase, which acts selectively only on the collagen fibers, provides gentle and bloodless optimum cleaning of the wounds of necrotic tissue. This enzyme isolated from the non-pathogenic strain of Streptomyces lavendulae (patent RU N 2075219, 10.03.97). The patent describes the collagenase isolated from the culture fluid of Streptomyces lavendulae strain with collagenolytic and total proteolytic activity that cleaves the beta chain of native collagen up to 84-86 kDa+26-KD has M m 110-120 kDa. In addition, the pharmaceutical composition is introduced Miramistin (FS 42-3498-98), which is a broad-spectrum antiseptic, not causing allergies. Further, the composition contains proxanol is chosen (NTU-6-14-164-87), with the ability to delay the exudate and pus from wounds.As the base composition includes a pharmaceutically acceptable glycol.Conducted comprehensive study of wound healing actions of the claimed composition: the study of the influence of composition on the rate of regeneration of skin damage in aseptic conditions, the impact on the speed at invizible the following adequate to the objectives of the experimental model.Model plane aseptic wounds of the skin.The model of linear wounds of the skin.Model infected (purulent) wounds.The model of burn wounds.To obtain the model and evaluation results used known methods. (Turishchev S. N., Pharmacology 4/5, 25-36, 1996)
Model plane aseptic wounds of the skin was obtained as follows.The experiments were conducted on white weinbrenner male mice weighing 20-30 g In animals under sterile conditions under anaesthesia (teminal sodium 50 mg/g) in the interscapular region of the back after first removing hair cut, a full-layer skin flap so that was oval wound area which was 80-100 mm For comparative study of dynamics and wound healing was performed visual observation and measurement of the area of RAS in 2-3 weeks daily, assessing the condition of the wound edges, its discharge, the formation of a scar.Treatment of animals was started immediately after surgery: wound inflicted the test substance in the form of ointment. Control animals received a placebo. As the comparison drug used is known for healing wounds - "Iruksol" (Mashkovsky). Speed healing of wounds received is R>< / BR>where d1- larger diameter of the oval;
d2- smaller diameter of the oval;
S - size of the wounds.Model linear skin wounds obtained in the experiment as follows.The experiments were conducted on white weinbrenner rats male weight of 300 g In animals under anesthesia (teminal sodium 40 mg/kg) in the interscapular region of the back after the preliminary hair removal inflicted 2 linear cut through all layers of the skin 1 cm long one on the right and left of the spine (approximately 2 cm from it). The wound was left one hub seam (sterile silk). For a comparative study of dynamics and wound healing was performed visual observation within 2-3 weeks daily, assessing the condition of the wound edges, its discharge, the formation of a scar. The quality of healing was evaluated by microscopic examination of the alignment. For this tissue from the defect area and adjacent to it the skin was fixed in a fluid Carnoy, embedded in paraffin and prepared slices (thickness of 8-10 μm. The sections were stained with hematoxylin and eosin.Treatment of animals was started immediately after surgery. The wound on the right put the test composition in the form of ointments. Left a wound of the same animal served as a control - it is ocena experimentally
In the experiments used white ainbridge rats weighing 300-320 g Under General anesthesia on the back neck area of the rats were vestigal hair and aseptically remove the skin from the subcutaneous tissue and superficial fascia. The wound was inserted Teflon ring (8 mm diameter) covered with the perforated tape. The edges and bottom of the advanced wound injured toothed clamp. In wound surface inside the ring were administered 0.5 ml of the suspension of the daily culture of pathogenic Staphylococcus aureus, strain 209 (1.5 billion microbial cells in 1 ml of physiologic R-RA). Rings were removed after 2 days and was performed by standard primary surgical processing of wounds, and then began treatment with the test substance. Control animals received placebo (materials).As methods for assessing the speed and completeness of healing used the following methods.Cytological examination of wound healing
Impression smears from the surface of the wound allows you to judge the nature of the wound and the cellular composition of the granulation. This smear is obtained by the touch of specially treated glass slides to the surface of the wound, pre-cleaned of pus. Drugs are air-dried, Phi is neutrophilic leukocytes, mononuclear cells, macrophages, fibroblasts, and their percentage.Microbiological examination of infected wounds
Microbiological control of efficiency of treatment of wounds is in the dynamics (every 3 days) compared with baseline (before treatment). Studied the qualitative composition of the microflora wounds and quantitative dynamics of microbial bodies in the wound.Histological studies in the dynamics of wound healing give an idea about qualitative changes in the wound under the influence of the drug. Used stained with hematoxilin and eosin
Morphometric study of scar tissue
Pieces of skin taken 13 days after surgery for histological examination were fixed in fluid Carnoy, manufactured stepped paraffin sections were stained with hematoxylin and eosin and viewed in the light microscope. Conducted a morphometric study of scar tissue: serial products using the grid Avtandilov gg method point account determined bulk density of the connective tissue and cellular elements. I calculated the percentage of fibroblasts to 500 cells.The results obtained for treatment is eskay, the significance of differences between groups was determined using the criterion Student.For comparative studies used 3 groups of animals.The first group of animals were treated with an ointment based on the declared composition, when the content of the active component 3 units, the second group of animals were treated with the ointment on the basis of the claimed compositions with a content of active component 7 units of the Third group of animals received treatment with the ointment "Iruksol". As a control in all three groups used a placebo.The results of the study are shown in tables 1 and 2.Table 1 shows the dynamics of wound healing back skin of mice (square planar RAS (ellipse) in mm2under the influence of the claimed composition.Table 2 shows the morphometric parameters characterizing the linear healing skin wounds in rats in the control and under the influence of the claimed composition.The results of studies carried out on models of linear and planar Russian Academy of Sciences, showed that the claimed composition has wound healing properties and, in addition, has a positive effect on the qualitative composition of scar tissue, Obrajuelo scar tissue RAS, treated with ointment.The invention is illustrated by the following examples of formulations.Example 1. Collagenase from Streptomyces lavendulae - 3 items
Miramistin - 0.1 g
Proxanol is chosen - 22,0 g
The glycol - the rest is up to 100 g
Example 2.Collagenase from Streptomyces lavendulae - 10 items
Miramistin - 0.5 g
Proxanol is chosen - 30,0 g
The glycol - the rest is up to 100 g
For Toxicological studies used active ingredient of the composition, i.e. containing enzymatic components, this ingredient has a brown color, odor, pH = 5,0-7,0.Study of acute toxicity of the substances was carried out on white mice (males and females weighing 19-21 g).The experiments were performed in accordance with the "guidelines for the study of General toxic action (drugs" (1997).The substance of the active ingredient was administered to the animals (mice) intravenously once daily, in the amount of 0.2-0.5 ml, with a speed of 0.1 ml/sThe duration of observation of the mice was 10-15 days. In the experiment, followed by behavior, appearance, motor activity, reaction of animals to external stimuli. In cases of death Phi and of the drug to mice in doses up to 3500 to 4000 U/kg signs of intoxication were observed. Animals remained agile, responding well to the environment. With further increase of the dose of the injected substance was noted the death of individual animals. Were installed srednetemperaturnyi dose (LD50for mice after a single intravenous substance: LD50=124002500 IU/kgTolerable upper intake of the substance when administered intravenously to mice at 4,500 U/kg In these cases, the observation of animals within 2 weeks of death is not marked.Research has also been undertaken on microbiological purity and progenote the claimed composition.Studies have been conducted on the presence of pyrogenic impurities in the drug. Testing was carried out on rabbits of the Chinchilla breed contained on a standard diet of the vivarium.As this drug is only applied externally as an ointment, to establish clear therapeutic dose difficult. Therefore, for the selection of the test-dose testing progenote we used the dose of collagenase drug - 5 IU/kgIt is established that the drug test dose of 5 IU per kg weight of the animal - pyrogenes.There was a trend of reduction of pyrogenic reactions with reducing woozy on progenote. Additional trials with new drug samples in a minimum volume of diluent, sound test dose may allow you to get the results, indicating a lack of progenote.A comparative study of antimicrobial activity of our songs and ointments "Iruksol" conducted by the "wells" (Methodical recommendations on experimental pre-clinical study of medicinal products for the topical treatment of wounds. - M., 1989), for which we used the environment N 3 and N 6 (formulation these environments is provided in these guidelines). As the test cultures were used freshly isolated clinical strains of microorganisms (S. aureus, S. epidermidis, E. Coli., Proteus sp., P. aeruginosa), from patients with purulent wounds of soft tissues.Isolation and identification of pure cultures was carried out by the method described in the methodological recommendations "Bacteriological diagnosis of wound infection" (M, 1984). The microorganisms were grown on solid nutrient medium on meat-peptone agar.Microbial suspension of the test cultures were prepared in accordance with the turbidity standard State Institute of sanitary control (GISC). L. A. Tarasevich. Concentration experienced what was Imerovigli in a thermostat at a temperature of - 37oC within 24 - 48 hours. The zone of growth inhibition of microorganisms accounted for the results.Studied antimicrobial activity:
- fundamentals of composition (combination of proxanol is chosen, poly (ethylene glycol);
- topical compositions without Miramistin (collagenase + proxanol is chosen + glycol);
- the claimed compositions in which the above basis, Miramistin and collagenase.The control drug was "Iruksol" is a combination drug, which is a fatty basis, collagenase and chloramphenicol.The results of the study of antimicrobial activity are shown in table 4.The results show that the developed composition ointment is an active drug against gram-positive and gram-negative flora. Given that the group Staphylococcus included coagulasenegative and metitsillinrezistentnye strains (25%), the results can be considered positive.Positive results against P. aeruginosa, the zone of growth inhibition of 15.8 mm indicates the sensitivity of Escherichia coli to test the ointment, although it is known that P. aeruginosa is often multidrug-resistant to antibacterial agents.the sensitivity can be compared as average, but it should be noted here that the components of the ointment base, referred to as a semi-permeable.According to modern concepts, the combination of polupronicaema and non-invasive components inside cells protects healthy tissue wounds from osmotic shock and makes using them harmless.As can be seen from the table, all the studied strains of microorganisms were resistant to ointments "Iruksol".Studies ointment for sterility showed absence of microorganisms in the claimed composition.Thus, studies have been conducted to study a new composition based on collagenase isolated from Streptomyces lavendulae, showed that the proposed new drug is a highly effective, non-toxic, non-allergenic tool for wound healing, burns and other skin damage. 1. Pharmaceutical composition based on collagenase microbial origin containing named enzyme, antiseptic ointment and a filler, characterized in that it contains collagenase from Streptomyces lavendulae at the rate of 3 - 10 IU per 100 g of product.2. The pharmaceutical composition under item 1, characterized in that it contains matemticas acceptable polyethylene glycol (PEG) in the following ratio of components:
Collagenase - 3 - 10 UNITS
Miramistin - 0.1 - 0.5 g
Proxanol is chosen - 22,0 - 30,0 g
PEG - the Rest is up to 100 g
< / BR>as a substrate for the determination of thrombin
FIELD: biotechnology, microbiology, medicine.
SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.
EFFECT: valuable medicinal properties of strain.
5 cl, 8 dwg, 10 ex
FIELD: biotechnology, microbiology, veterinary science.
SUBSTANCE: for preparing a preparation cells of microorganism Escherichia coli VKM CR-322D is cultured in nutrient medium containing Hottinger's broth, glucose, yeast extract, manganese sulfate, potassium hydrophosphate, sodium chloride and tap water in the content of amine nitrogen 125-155 mg%. Glucose is added by batch portions in the process of culturing cells that is carried out at temperature 30-31oC at stirring and aeration for 10-12 h. Prepared cultural fluid containing 3 x 109 bacterial cells/ml is mixed with protective sucrose-gelatin medium and subjected for lyophilic drying. Dried mass is stored under nitrogen that enhances safety of viable cells in the preparation. Applying the preparation for prophylaxis and treatment of agricultural and domestic animals and poultries with gastroenteric diseases provides its effectiveness.
EFFECT: improved preparing method, valuable veterinary properties of preparation.
17 cl, 8 ex
FIELD: biotechnology, food and medicinal industry, microbiology.
SUBSTANCE: the strain Bifidobacterium longum 379-IN is obtained by selection without using methods of genetic modification of the strain Bifidobacterium longum B379M and distinct by ability to utilize insulin. The strain is deposited in GKNM GU "MNIIEM named for G. N. Gabrichevskiy Russia Ministry of Public Health" at № 172. The strain shows high technological effectiveness, accumulates biomass with substrates of vegetable origin and artificial nutrient media for short periods with concentration of bifidobacteria, it elicits acid-forming and antagonistic properties with respect to pathogenic and putrid microflora. This allows its using in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, ferments, hygienic and cosmetic agents providing probiotic effect and normalization of microbiocenosis in human body, among them in gastroenteric and urogenital tracts, cutaneous and mucosa integuments. Invention can be used in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, hygienic and cosmetic agents.
EFFECT: valuable properties of strain, expanded assortment of similar agents.
FIELD: biotechnology, medicine, infectious diseases, medicinal microbiology.
SUBSTANCE: invention relates to a composition designated for treatment and prophylaxis of infections caused by Neisseria microorganism that comprises the following components: (a) protein with amino acid sequence similar by 65% and above with the natural Neisseria protein of a single species (the first group of amino acid sequences is given in the text) and/or its fragment consisting of 10 and more amino acids and eliciting antigen properties; (b) the second protein with amino acid sequence similar by 65% and above with the natural Neisseria protein of another species (the second group of amino acid sequences with even numbers is given in the text), and/or its fragment consisting of 10 or more amino acids and eliciting antigen properties; in particular, the second protein represents NspA. The composition comprises additionally adjuvant. The composition is used both a medicinal agent and for manufacturing the medicinal agent. Applying the invention provides enhancing the effectiveness of prophylaxis or treatment due to the universal effect of the composition (vaccine). Invention can be used in medicine for treatment of infections.
EFFECT: valuable medicinal properties of composition.
8 cl, 137 dwg, 5 tbl, 12 ex
FIELD: biotechnology, medicinal microbiology.
SUBSTANCE: invention relates, in particular, to a method for preparing nutrient medium used for production of bacteriophages. Nutrient medium for production of bacteriophages contains acidic casein hydrolyzate as a base with hydrolysis degree 0.6-0.7 and vitamins: nicotinic acid, folic acid, calcium pantothenate, riboflavin, thiamine bromide and biotin, and distilled water. Also, invention relates to a method for preparing nutrient medium used for production of bacteriophages involving hydrolysis of natural casein mixed with water using hydrochloric acid under pressure 0.2 ± 0.05 MPa up to attainment of hydrolysis degree 0.6-0.7 being isoprecipitation and treatment with carbon are carried out simultaneously. Invention provide enhancing growth properties of medium and quality of bacteriophages produced and reducing the cost.
EFFECT: improved preparing method.
3 cl, 1 ex